Fig 2: PMAIP1 deficiency reduces heat-induced death and delays caspase-3 and caspase-7 activation.a Apoptosis and b necrosis of PMAIP1(+) and PMAIP1(−) HECa10 cells monitored between 2 and 24 h after heat shock or during bortezomib (Bort) or camptothecin (CPT) treatments. Shown are mean values ± SD from one (representative) of three independent experiments; the statistically significant difference between treated and untreated samples or PMAIP1(+) and PMAIP1(−) samples is marked with an asterisk (*p < 0.05, **p < 0.001). c PMAIP1(+) and PMAIP1(−) cells were heat-shocked for 1 h at 43 °C and protein extracts were analyzed by western blot up to 6 h of recovery. ACTB was used as a loading control. C untreated cells, CPT camptothecin treatment of wild-type HECa10 cells for 6 h (positive control for PMAIP1 induction); unspecific protein band recognized by anti-PMAIP1 Ab is marked with an asterisk. The graphs show the results of densitometric analyses from three independent experiments; *p < 0.05, **p < 0.001.

Fig 3: Antitumor activity of chidamide (CHI) and/or doxorubicin (DOX) in MDA breast cancer cells in vivo. (A–C) CALDOX xenograft tumor growth curve, size, and body weight after treatment with normal saline (control), DOX, CHI or CHI + DOX. (D) Western blot analysis of HDAC1, H3K9, H3K18, p53, p21, Puma, Bcl-xl, Bcl-2, cleaved caspase-7, cleaved caspase-3, and Bax on CALDOX-derived tumors treated with PBS (control), DOX, CHI, or CHI + DOX. (E) TUNEL staining analyzed cell apoptosis after treatment with normal saline (control), DOX, CHI, or CHI + DOX. (F) Relative fold change of Bax, Bcl-2, and caspase-3 gene expression levels in CALDOX-derived tumors treated with normal saline (control), DOX, CHI, or CHI + DOX. The numerical values are showed as mean ± SD of three independent replicates. “*” indicates a significant difference compared with the control group (*P < 0.05, **P < 0.01,***P<0.001),”#” indicates a significant difference compared with the DOX-treated group (##P < 0.01, ###P < 0.001), and “&” indicates a significant difference compared with the CHI-treated group (&P < 0.05, &&P < 0.01). ns, no significance.

Fig 4: Increased cleavage of PARP and caspases in TAT‐treated cells. (A) Western blot of PARP and its cleavage products in HeLa cells treated with or without 25 μm TAT for each indicated time. Staurosporine was used as the positive control and tubulin as the loading control. (B) Western blots of caspase‐7 and caspase‐9 cleavage products after treatment of HeLa cells with 25 or 50 μm TAT for 48 h.

Fig 5: Autophagic response of sorafenib-resistant cells to sorafenib or lenvatinib. (A) Huh-7SR and Hep-3BSR cells were treated with 10 μM sorafenib or lenvatinib for 6 h. LC3, p62, caspase-7, caspase-3, and PARP expression were analyzed using Western blot. C.: control; Sor: sorafenib; Len: lenvatinib. (B) A lower p62 level is associated with longer overall survival of HCC patients based on TCGA database analysis (http://www.oncolnc.org, accessed on 5 October 2021).